三叶青乙酸乙酯提取物诱导人结肠癌HT-29细胞凋亡

Induction of Apoptosis by Ethyl-acetate Fraction of Extracts from Tetrastigma Hemsleyanum Diels et.Gilg in Human Colon Cancer HT-29 Cells

目的:评价三叶青乙酸乙酯提取物(ethyl-acetate fraction of extracts from Tetrastigma hemsleyanumDiels et.Gilg,EAFT)诱导人结肠癌HT-29细胞凋亡作用。方法:体外培养HT-29细胞。FITC标记annexinⅤ/PI染色流式细胞术分析细胞凋亡率。ELISA法测定细胞组蛋白/DNA碎片。Western Blotting分析细胞Bax蛋白和细胞色素C的表达。结果:EAFT以浓度依赖方式诱导annexinⅤ染色阳性细胞百分率和组蛋白/DNA碎片增加(P<0.01)。10.0 mg/L EAFT分别作用HeLa细胞6 h、12 h和24 h,细胞色素C和Bax蛋白表达上调。结论:EAFT具有诱导人结肠癌HT-29细胞凋亡作用,其作用机制与激活线粒体途径有关。

Objective To evaluate the effects of ethyl-acetate fraction of extracts from Tetrastigma hemsleyanum Diels et.Gilg(EAFT) on the apoptotic cell death in human colon cancer HT-29 cell line.Methods Human colon cancer HT-29 cells were cultured in vitro.Flow cytometry after FITC labeled annexin V staining was used to determine the percentage of apoptotic cells.Histone/DNA fragments were determined using ELISA assay.Western Blotting was used to analyze the expresson of Bax and cyto C protein in HT-29 cells.Results EAFT significantly induced increase of the percentage of apoptotic cells(P<0.05),and increased leakage of the histone/DNA fragments in HT-29 cell line(P<0.05),in a concentration-dependent manner.The upregulation of the expression level of Bax and cyto c protein in HT-29 cells treated with EAFT at 10mg/L for 6,12,24 h(P<0.05),as assessment by Western blotting.Conclusion EAFT can effectively induce apoptosis of human colon cancer HT-29 cells,and its mechanism is involved in activation of mitochondrial apoptotic pathway.